Multiplex nested polymerase chain reaction for Treponema pallidum using blood is more sensitive than using serum

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Ida Effendi
Yeva Rosana
Andi Yasmon
Wresti Indriatmi


Syphilis is a multistage disease transmitted primarily through sexual intercourse. Nowadays, the polymerase chain reaction (PCR) test for Treponema pallidum has been widely used and is expected to overcome problems in diagnostic tests for syphilis. The Treponema pallidum PCR is influenced by type of specimens, PCR methods and target genes. This study aimed to assess the use of blood and serum in multiplex nested PCR for Treponema pallidum, targeting the 23S rRNA.

A cross-sectional study was conducted from April 2015 - April 2016. Sampling was carried out consecutively among patients with clinical features of secondary syphilis who came to Sexually Transmitted Disease (STD) clinics in Jakarta. All sera were also tested with Rapid Plasma Reagin (RPR) and Treponema pallidum Hemagglutination Assay (TPHA) assay, which was considered as the gold standard for this study. We determined the sensitivity and specificity of the multiplex nested PCR for Treponema pallidum using blood and serum.

PCR test was performed on 122 clinical specimens (61 blood and 61 serum). The positive results of PCR test on blood was 22.95% and serum was 6.56%, while the positive results of serology was 68.85%. The sensitivity of Treponema pallidum multiplex nested PCR on blood was 30.95% compared to serum 9.52% (p=0.006). PCR test on blood is able to detect 3.25 times higher than serum.

The use of blood has a higher proportion of positives compared to serum in Treponema pallidum multiplex nested PCR using 23S rRNA target gene.

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How to Cite
Effendi, I., Rosana, Y., Yasmon, A., & Indriatmi, W. (2018). Multiplex nested polymerase chain reaction for Treponema pallidum using blood is more sensitive than using serum. Universa Medicina, 37(1), 75–84.
Original Articles


Wijesooriya NS, Rochat RW, Kamb ML, et al. Global burden of maternal and congenital syphilis in 2008 and 2012: a health systems modelling study. Lancet Glob Health 2016;4:e525–33.

Braxton J, Carey D, Davis D, et al. Sexually transmitted disease surveillance 2014. Atlanta, Georgia: U.S. Department of Health and Human Services , Centers for Disease Control and Prevention National Center for HIV/AIDS, Viral Hepatitis, STD, and TB Prevention Division of STD Prevention; 2015.

Kementerian Kesehatan Republik Indonesia. Surveilans terpadu biologi perilaku. Jakarta: Kementerian Kesehatan Republik Indonesia Dirjen Pengendalian Penyakit dan Penyehatan Lingkungan; 2011.

World Health Organization. Global incidence and prevalence of selected curable sexually transmitted infections-2008. Geneva: World Health Organization;2012.

World Health Organization. Report on global sexually transmitted infection surveillance 2013. Geneva: World Health Organization;2014.

Tramont EC. Treponema pallidum (syphilis). In: Bennett JE, Dolin R, Blaser MJ, editors. Mandell, Douglas, and Bennett’s principles and practice of infectious diseases. Philadelphia: Elsevier Saunders;2015.p.3035-55.

Zobaníkova M, Mikolka P, Èejková D, et al. Complete genome sequence of Treponema pallidum strain DAL-1. Standards Genomic Sci 2012;7:12-21. DOI:10.4056/sigs.2615838.

Tipple C, Hanna MOF, Hill S, et al. Getting the measure of syphilis: qPCR to better understand early infection. Sex Transm Infect 2011;87:e485. doi: 10.1136/sti.2011.049494.

.9 Gayet-Ageron A, Combescure C, Lautenschlager S, et al. Comparison of the diagnostic accuracy of polymerase chain reaction targeting the 47kDa protein membrane gene of Treponema pallidum or the DNA polymerase I gene: a systematic review and meta-analysis. J Clin Microbiol 2015; 53:3522-9. doi: 10.1128/JCM.01619-15.

Grange PA, Gressier L, Dion PL, et al. Evaluation of a PCR test for detection of Treponema pallidum in swabs and blood. J Clin Microbiol 2012;50:546-52. doi: 10.1128/JCM.00702-11.

Casal CAD, Silva MOD, Costa IB, et al. Molecular detection of Treponema pallidum sp. pallidum in blood samples of VDRL-seroreactive women with lethal pregnancy outcomes: a retrospective observational study in northern Brazil. Revista da Sociedade Brasileira de Medicina Tropical 2011;44:451-6.

Buffet M, Grange PA, Gerhardt P, et al. Diagnosing Treponema pallidum in secondary syphilis by PCR and immunohistochemistry. J Invest Dermatol 2007;127:2345–50. doi: 10.1038/sj.jid.5700888.

Cruz AR, Pillay A, Zuluaga AV, et al. Secondary syphilis in Cali, Colombia: new concepts in disease pathogenesis. PLoS Negl Trop Dis 2010;4:e690. doi: 10.1371/journal.pntd.0000690.

Ho EL, Lukehart SA. Syphilis: using modern approaches to understand an old disease. J Clin Invest 2011;121:4584-92.

Gayet-Ageron A, Toutus-Trelllu NBL, Lautenschlager S, et al. Assessment of a real-time PCR test to diagnose syphilis from diverse biological samples. Sex Transm Infect 2009;85:264-9. doi: 10.1136/sti.2008.034314.

Gultom DA, Rosana Y, Efendi I, et al. Detection and identification of azithromycin resistance mutations on Treponema pallidum 23S rRNA gene by nested multiplex polymerase chain reaction. Med J Indones 2017;26:90-6.

Sharma S, Chaudhary J, Hans C. VDRL v/s TPHA for diagnosis of syphilis among HIV sero-reactive patients in a tertiary care hospital. Int J Curr Microbiol App Sci 2014;3:726-30.

Martin IE, Tsang RSW, Sutherland K, et al. Molecular characterization of syphilis in patients in Canada: azithromycin resistance and detection of Treponema pallidum DNA in whole-blood samples versus ulcerative swabs. J Clin Microbiol 2009;47:1668-73.

Tipple C, Jones R, McClure M, et al. Rapid Treponema pallidum clearance from blood and ulcer samples following single dose benzathine penicillin treatment of early syphilis. PLoS Negl Trop Dis 2015;10:e0003492. doi: 10.1371/ journal. pntd.0003492.

Gayet-Ageron A, Lautenschlager S, Ninet B, et al. Sensitivity, specificity and likelihood ratios of PCR in the diagnosis of syphilis: a systematic review and meta-analysis. Sex Transm Infect 2013;89:251-6.

Sato NS. Laboratorial diagnosis of syphilis. Syphilis - recognition, description and diagnosis. Croatia: INTECH;2011.p.87-108.

Wu BR, Tsai MS, Yang CJ, et al. Spirochetemia due to Treponema pallidum using polymerase-chain-reaction assays in patients with early syphilis: prevalence, associated factors and treatment response. Clin Microbiol Infect 2014;20:O524-7. doi: 10.1111/1469-0691.12504.

Leslie DE, Azzato F, Karapanagiotidis T, et al. Development of a real-time PCR assay to detect Treponema pallidum in clinical sspecimens and assessment of the assay’s performance by comparison with serological testing. J Clin Microbiol 200745:93-6. doi:10.1128/JCM.01578-06.